In vitro culture and biological characteristics of human bone marrowª²derived endothelial progenitor cells

    ZHENG Qiª²Jun, WANG Ruiª²An, CAI Zhenª²Jie, YU Shiª²Qiang, OUYANG Hui, ZHAO Jianª²Bin

    1Department of Cardiovascular Surgery, Xijing Hospital, Fourth Military Medical University, Xi¬ðan 710033, China, 2Department of Anesthesiology, Changan Hospital of Xi¬ðan, Xi¬ðan 710016, China

    ¡¾Abstract¡¿ AIM: To purify and culture human bone marrowª²derived endothelial progenitor cells (EPCs) in vitro and to observe their biological characteristics. METHODS: Mononuclear cells were isolated from bone marrow suspension by density gradient centrifugation and EPCs were harvested in the endothelial growth medium (EGMª²2). EPCs were observed under microscope, the cell growth was calculated by cell counting and the cells were identified by electronic microscope examination and immunofluorescence staining for the expression of ¢ø/vWF, vascular endothelial growth factor receptorª²2 (VEGFRª²2). RESULTS: The cultureª²dishª²adherent EPCs uniformly had a round or spindle shape and reformed clones with cobbleª²stone morphology. The cells could be identified by the positive staining for VEGFRª²2 and ¢ø/vWF. Weibelª²Paladle bodies could be seen under transmission electron microscope. CONCLUSION: EPCs are one group of stem cells in the bone marrow with the capacity of differentiating into endothelial cells. They can be purified and cultured by density gradient centrifugation and subjected to culture in vitro with endothelial growth medium (EGMª²2 MV). These findings suggest that EPCs may be a new seed cell of tissue engineering. ......